Validation of an assay for quantification of alpha-amylase in saliva of sheep
The objective of this study was to develop a time-resolved immunofluorometric assay (TR-IFMA) for quantification of salivary alpha-amylase in sheep. For that purpose, after the design of the assay, an analytical and a clinical validation were carried out. The analytical validation of
the assay showed intra- and inter-assay coefficients of variation (CVs) of 6.1% and 10.57%, respectively and an analytical limit of detection of 0.09 ng/mL. The assay also demonstrated a high level of accuracy, as determined by linearity under dilution. For clinical validation, a model of
acute stress testing was conducted to determine whether expected significant changes in alpha-amylase were picked up in the newly developed assay. In that model, 11 sheep were immobilized and confronted with a sheepdog to induce stress. Saliva samples were obtained before stress induction
and 15, 30, and 60 min afterwards. Salivary cortisol was measured as a reference of stress level. The results of TR-IFMA showed a significant increase (P < 0.01) in the concentration of alpha-amylase in saliva after stress induction. The assay developed in this study could be used
to measure salivary alpha-amylase in the saliva of sheep and this enzyme could be a possible noninvasive biomarker of stress in sheep.
Document Type: Research Article
Publication date: 01 July 2016
- The Canadian Journal of Veterinary Research (CJVR), published by Canadian Veterinary Medical Association, is Canada's only veterinary research publication. This quarterly peer-reviewed journal has earned a wide international readership through the publishing of high quality scientific papers in the field of veterinary medicine. CJVR publishes the results of original research in veterinary and comparative medicine.
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